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Development of an Improved Loop-Mediated Isothermal Amplification Assay for On-Site Diagnosis of Fire Blight in Apple and Pear

  • Shin, Doo-San;Heo, Gwang-Il;Son, Soo-Hyeong;Oh, Chang-Sik;Lee, Young-Kee;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.34 no.3
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    • pp.191-198
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    • 2018
  • Fast and accurate diagnosis is needed to eradicate and manage economically important and invasive diseases like fire blight. Loop-mediated isothermal amplification (LAMP) is known as the best on-site diagnostic, because it is fast, highly specific to a target, and less sensitive to inhibitors in samples. In this study, LAMP assay that gives more consistent results for on-site diagnosis of fire blight than the previous developed LAMP assays was developed. Primers for new LAMP assay (named as DS-LAMP) were designed from a histidine-tRNA ligase gene (EAMY_RS32025) of E. amylovora CFBP1430 genome. The DS-LAMP amplified DNA (positive detection) only from genomic DNA of E. amylovora strains, not from either E. pyrifoliae (causing black shoot blight) or from Pseudomonas syringae pv. syringae (causing shoot blight on apple trees). The detection limit of DS-LAMP was 10 cells per LAMP reaction, equivalent to $10^4$ cells per ml of the sample extract. DS-LAMP successfully diagnosed the pathogens on four fire-blight infected apple and pear orchards. In addition, it could distinguish black shoot blight from fire blight. The $B{\ddot{u}}hlmann$-LAMP, developed previously for on-site diagnosis of fire blight, did not give consistent results for specificity to E. amylovora and on-site diagnosis; it gave positive reactions to three strains of E. pyrifoliae and two strains of P. syringae pv. syringae. It also, gave positive reactions to some healthy sample extracts. DS-LAMP, developed in this study, would give more accurate on-site diagnosis of fire blight, especially in the Republic of Korea, where fire blight and black shoot blight coexist.

Amperometric Biosensor for Hydrogen Peroxide Determination Based on Black Goat Liver-Tissue and Ferrocene Mediation (흑염소 간-조직과 Ferrocene 매개체를 사용한 과산화수소정량 전류법 바이오센서)

  • Kwon, Hyo-Shik;Park, In-Keun;Kim, Yang-Sug
    • Journal of the Korean Chemical Society
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    • v.48 no.5
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    • pp.491-498
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    • 2004
  • The response characteristics of the bioelectrode developed by the co-immobilization of black goat liver tissue and ferrocene in a carbon paste matrix for the amperometric determination of hydrogen peroxide were evaluated. In the range of electrode potential examined ($-0.3{\sim}+0.0\;V$ vs. Ag/AgCl), the response time was relatively short (t95%=12 s) and it responded in the wide range of pH. The detection limit was 2.25${\times)10^{-6}M$ and a relative standard deviation of the measurements which were repeated 15 times using 1.0${\times}10^{-2 }$M hydrogen peroxide was 1.87%. The bioelectrode sensitivity decreased to 50% of the original value in 19 days of continuous use.

Prediction of Depression among Elderly with Mild Cognitive Impairment Living in the Community (경도인지장애 노인의 우울 예측 요인)

  • Shin, Kyung-Rim;Kang, Youn-Hee;Kim, Mi-Young;Jung, Duk-Yoo;Lee, Eliza
    • Korean Journal of Adult Nursing
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    • v.24 no.2
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    • pp.171-182
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    • 2012
  • Purpose: This study aimed to identify depression and its predictors among Korean community-dwelling elderly with mild cognitive impairment. Methods: Secondary data analyses of the data collected by the "Study on tailored integration program for reinforcing cognitive and physical function of the frail elderly. The study used data from one-to-one interviews using structured questionnaires. The subjects were 346 community-dwelling elderly who visited a healthcare center in Seoul, Korea. A descriptive correlational study design was utilized to explore depression and its predictors including physical factors and psychosocial factors among the elderly with mild cognitive impairment. Data were analyzed using t-test, Pearson's correlation, and hierarchical regression. Results: Mean score of depression was within normal limit $11.61{\pm}6.69$. Somatic symptoms, negative life events (severity), social support were shown as significant predictors of depression. The most influential predictor for depression was somatic symptoms (${\beta}$=.340, $p$ <.001). Conclusion: To prevent and relieve depression in the elderly with mild cognitive impairment, nursing intervention strategies which consider well-balanced physical and psychosocial aspects are needed. In particular, a specific nursing intervention strategy is required to improve physical health of the elderly with mild cognitive impairment.

A Case Study on the Development and Evaluation of an Web-based Learning Program (웹기반 교육 프로그램의 개발과 프로그램 운영에 따른 효과 고찰)

  • 이영미;장정옥;오유진
    • Journal of Nutrition and Health
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    • v.35 no.8
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    • pp.886-895
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    • 2002
  • Introduction and application of virtual education has been rapidly increased in these days. A variation of information communication technology has an effect on education in interconnect with network as internet in the world that exceed the limit of time and regional. Computer and network communication technology through the medium of internet make an entrance cyber education as a new education paradigm. It must be affective on learner who have various educational characteristics and requirements. It begins to appear quality, quantity improvements of knowledge and the development of information technology that web based cyber education. This study was conducted to develop the web based education program and to evaluate the effectiveness of learning satisfaction and accomplishment and to compare the cyber lecture system with the traditional lecture system During the second semester of 2001, this study was investigated 317 registered students in a "Food and Culture" class at Kyungwon University. The data were obtained from pre and post-study with self-administered questionnaire. The evaluation and satisfaction score of students who were registered in cyberclass was negative tendency to compare pre with post-test scores, because of insuffciency of computer-aided lecture system. The major problem was inconvenient in checking system for connecting times in cyberclass which was one of evaluation point in final score. Another problem was frequently disconnection during cyber studying and not to concentrate each time in the cyber lecture because of eye fatigue, boring due to less interesting contents than other newly developed web-site. The students was prefer to mix type of the cyber and traditional lecture type class. The result of final score an each class, the score of cyber class (71.36 $\pm$ 22.44) was significantly lower than other groups (mixed type : 76.66 $\pm$ 19.99, traditional type :79.17 $\pm$ 15.72) (p < 0.05). Cyber class was attempted to present a useful and interesting teaching and learning tool which can be applied successfully in a longer term. The result suggest that various teaching and learning strategies should be developed considering the fact that the student learn alone most in time.t in time.

Evaluation of stability of interface between CCM (Co-Cr-Mo) UCLA abutment and external hex implant

  • Yoon, Ki-Joon;Park, Young-Bum;Choi, Hyunmin;Cho, Youngsung;Lee, Jae-Hoon;Lee, Keun-Woo
    • The Journal of Advanced Prosthodontics
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    • v.8 no.6
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    • pp.465-471
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    • 2016
  • PURPOSE. The purpose of this study is to evaluate the stability of interface between Co-Cr-Mo (CCM) UCLA abutment and external hex implant. MATERIALS AND METHODS. Sixteen external hex implant fixtures were assigned to two groups (CCM and Gold group) and were embedded in molds using clear acrylic resin. Screw-retained prostheses were constructed using CCM UCLA abutment and Gold UCLA abutment. The external implant fixture and screw-retained prostheses were connected using abutment screws. After the abutments were tightened to 30 Ncm torque, 5 kg thermocyclic functional loading was applied by chewing simulator. A target of $1.0{\times}10^6$cycles was applied. After cyclic loading, removal torque values were recorded using a driving torque tester, and the interface between implant fixture and abutment was evaluated by scanning electronic microscope (SEM). The means and standard deviations (SD) between the CCM and Gold groups were analyzed with independent t-test at the significance level of 0.05. RESULTS. Fractures of crowns, abutments, abutment screws, and fixtures and loosening of abutment screws were not observed after thermocyclic loading. There were no statistically significant differences at the recorded removal torque values between CCM and Gold groups (P>.05). SEM analysis revealed that remarkable wear patterns were observed at the abutment interface only for Gold UCLA abutments. Those patterns were not observed for other specimens. CONCLUSION. Within the limit of this study, CCM UCLA abutment has no statistically significant difference in the stability of interface with external hex implant, compared with Gold UCLA abutment.

A Solid-Contact Indium(III) Sensor based on a Thiosulfinate Ionophore Derived from Omeprazole

  • Abbas, Mohammad Nooredeen;Amer, Hend Samy
    • Bulletin of the Korean Chemical Society
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    • v.34 no.4
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    • pp.1153-1159
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    • 2013
  • A novel solid-contact indium(III)-selective sensor based on bis-(1H-benzimidazole-5-methoxy-2-[(4-methoxy-3, 5-dimethyl-1-pyridinyl) 2-methyl]) thiosulfinate, known as an omeprazole dimer (OD) and a neutral ionophore, was constructed, and its performance characteristics were evaluated. The sensor was prepared by applying a membrane cocktail containing the ionophore to a graphite rod pre-coated with polyethylene dioxythiophene (PEDOT) conducting polymer as the ion-to-electron transducer. The membrane contained 3.6% OD, 2.3% oleic acid (OA) and 62% dioctyl phthalate (DOP) as the solvent mediator in PVC and produced a good potentiometric response to indium(III) ions with a Nernstian slope of 19.09 mV/decade. The constructed sensor possessed a linear concentration range from $3{\times}10^{-7}$ to $1{\times}10^{-2}$ M and a lower detection limit (LDL) of $1{\times}10^{-7}$ M indium(III) over a pH range of 4.0-7.0. It also displayed a fast response time and good selectivity for indium(III) over several other ions. The sensor can be used for longer than three months without any considerable divergence in potential. The sensor was utilized for direct and flow injection potentiometric (FIP) determination of indium(III) in alloys. The parameters that control the flow injection method were optimized. Indium(III) was quantitatively recovered, and the results agreed with those obtained using atomic absorption spectrophotometry, as confirmed by the f and t values. The sensor was also utilized as an indicator electrode for the potentiometric titration of fluoride in the presence of chloride, bromide, iodide and thiocyanate ions using indium(III) nitrate as the titrant.

Amperometric Detection of Some Catechol Derivatives and o-aminophenol Derivative with Laccase Immobilized Electrode: Effect of Substrate Structure

  • Quan De;Shin Woonsup
    • Journal of the Korean Electrochemical Society
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    • v.7 no.2
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    • pp.83-88
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    • 2004
  • [ $DeniLite^{TM}$ ] laccase immobilized Pt electrode was used for amperometric detection of some catechol derivatives and o-aminophenol (OAP) derivative by means of substrate recycling. In case of catechol derivatives, the obtained sensitivities are 85, 79 and $57 nA/{\mu}M$ with linear ranges of $0.6\~30,\;0.6\~30\;and\; 1\~25 {\mu}M$ and detection limits (S/N=3) of 0.2, 0.2 and $0.3{\mu}M$ for 3,4-dihydroxycinnaminic acid (3,4-DHCA), 3,4-dihydroxybenzoic acid (3,4-DHBA) and 3,4-dihydroxyphenylacetic acid (3,4-DHPAA), respectively. In case of OAP derivative, the obtained sensitivity is $237 nA/{\mu}M$ with linear range of $0.2\~15{\mu}M$ and detection limit of 70 nM for 2-amino-4-chlorophenol (2-A-4-CP). The response time $(t_{90\%})$ is about 2 seconds for each substrate and the long-term stability is around 40-50days for catechol derivatives and 30 days for 2-A-4-CP with retaining $80\%$ of initial activity. The optimal pHs of the sensor for these substrates are in the range of 4.5-5.0, which indicates that stability of the enzymatically oxidized product plays a very important role in substrate recycling. The different sensitivity of the sensor for each substrate can be explained by the electronic effect of the sugstituent on the enzymatically oxidized form.

Kinetic Study on Aminolysis of 4-Chloro-2-Nitrophenyl X-Substituted-Benzoates in Acetonitrile and in 80 mol % H2O/20 mol % DMSO: Effect of Medium on Reactivity and Reaction Mechanism

  • Kim, Ha-Ram;Um, Tae-Il;Kim, Min-Young;Um, Ik-Hwan
    • Bulletin of the Korean Chemical Society
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    • v.35 no.4
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    • pp.1128-1132
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    • 2014
  • A kinetic study on aminolysis of 4-chloro-2-nitrophenyl X-substituted-benzoates (6a-i) in MeCN is reported. The Hammett plot for the reactions of 6a-i with piperidine consists of two intersecting straight lines, while the Yukawa-Tsuno plot exhibits an excellent linear correlation with ${\rho}_X$ = 1.03 and r = 0.78. The nonlinear Hammett plot is not due to a change in rate-determining step (RDS) but is caused by the resonance stabilization of substrates possessing an electron-donating group in the benzoyl moiety. The Br${\phi}$nsted-type plot for the reactions of 4-chloro-2-nitrophenyl benzoate (6e) with a series of cyclic secondary amines is linear with ${\beta}_{nuc}$ = 0.69, an upper limit for reactions reported to proceed through a concerted mechanism. The aminolysis of 6e in aqueous medium has previously been reported to proceed through a stepwise mechanism with a change in RDS on the basis of a curved Br${\phi}$nsted-type plot. It has been concluded that instability of the zwitterionic tetrahedral intermediate ($T^{\pm}$) in MeCN forces the reaction to proceed through a concerted mechanism. This is further supported by the kinetic result that the amines used in this study are less reactive in MeCN than in $H_2O$, although they are more basic in MeCN over 7 $pK_a$ units.

Qualitative and quantitative assessment of process related impurities in Brigatinib raw material and formulations using HPLC

  • Attada Tharun;Potnuru Jagadeesh;B Srinivasa Kumar;Kota Thirumala Prasad;Venkateswara Rao Anna
    • Analytical Science and Technology
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    • v.36 no.4
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    • pp.180-190
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    • 2023
  • The presence of process related impurities in any drug or the drug product was associated with its safety, stability and efficacy. The overall literature survey proved that there is no method published on the assessment of process related impurities in brigatinib. In this study, a simple, reliable and stable HPLC qualitative method was reported for quantification of process related impurities with easy and quick extraction procedure. The impurities along with standard brigatinib was resolved on Lichrospher® C18 (250 mm × 4.6 mm; 5 ㎛ particle size) column in room temperature using methanol, acetonitrile, pH 4.5 phosphate buffer in 55:25:20 (v/v) at 1.0 mL/min as mobile phase and UV detection at 261 nm. The method produces well resolved peaks at retention time of 4.60 min, 12.28 min, 3.37 min, 7.34 min and 8.39 min respectively for brigatinib, impurity A, B, C and D. The method produces a very sensitive detection limit of 0.0065 ㎍/mL, 0.0068 ㎍/mL, 0.0053 ㎍/mL and 0.0058 ㎍/mL for impurity A, B, C and D respectively with calibration curve linear in the concentration range of 22.5-135 ㎍/mL for brigatinib and 0.0225-0.135 ㎍/mL for impurities. The method produces all the validation parameters under the acceptable level and doesn't produces any considerable changes in peak area response while minor changes in the developed method conditions. The method can effectively resolve the unknown stress degradation products along with known impurities with less % degradation. The method can efficiently resolve and quantify the impurities in formulation and hence can suitable for the routine quality analysis of brigatinib in raw material and formulation.

A Study on Comparison of Response Time using Open API of Daishin Securities Co. and eBestInvestment and Securities Co.

  • Ryu, Gui Yeol
    • International journal of advanced smart convergence
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    • v.11 no.1
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    • pp.11-18
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    • 2022
  • Securities and investment services have and use large data. Investors started to invest through their own analysis methods. There are 22 major securities and investment companies in Korea and only 6 companies support open API. Python is effective for requesting and receiving, analyzing text data from open API. Daishin Securities Co. is the only open API that officially supports Python, and eBest Investment & Securities Co. unofficially supports Python. There are two important differences between CYBOS plus of Daishin Securities Co. and xingAPI of eBest Investment & Securities Co. First, we must log in to CYBOS plus to access the server of Daishin Securities Co. And the python program does not require a logon. However, to receive data using xingAPI, users log on in an individual Python program. Second, CYBOS plus receives data in a Request/Reply method, and zingAPI receives data through events. It can be thought that these points will show a difference in response time. Response time is important to users who use open APIs. Data were measured from August 5, 2021, to February 3, 2022. For each measurement, 15 repeated measurements were taken to obtain 420 measurements. To increase the accuracy of the study, both APIs were measured alternately under same conditions. A paired t-test was performed to test the hypothesis that the null hypothesis is there was no difference in means. The p-value is 0.2961, we do not reject null hypothesis. Therefore, we can see that there is no significant difference between means. From the boxplot, we can see that the distribution of the response time of eBest is more spread out than that of Cybos, and the position of the center is slightly lower. CYBOS plus has no restrictions on Python programming, but xingAPI has some limits because it indirectly supports Python programming. For example, there is a limit to receiving more than one current price.